Method for solid-phase synthesis of DNA encoded compound library

Abstract

The present invention provides a method of solid-phase synthesis of DNA-encoded compound library. The method includes following steps: a) reacting solid carrier G-1 with linker molecule L-1 to prepare L-G-1; b) reacting DNA with linker molecule L-0 to prepare L-2; c) reacting L-G-1 with L-2 to prepare L-G-2; d) removing protection group of the L-G-2 and obtaining L-G-2-1; e) reacting the L-G-2-1 with synthetic building block and performing DNA encoding; and f) removing the solid carrier and obtaining the DNA-encoded compound library. Compared with the prior art, the present invention can complete post-treatment purification of the reaction only by filtration and irrigation processes for several times. The present invention is simple to operate, can shorten the production cycle of DNA encoded compound library with more than 50%, significantly increases the production efficiency and the unicity as well as the purity of the final products.

What is claimed is: 
     
       1. A method for solid-phase synthesis of a DNA-encoded compound library, comprising:
 (a) reacting controlled pore glass (CPG) beads comprising a free amine as a solid carrier with a first linker 
 
       
         
           
           
               
               
           
         
       
       in an organic solvent to obtain 
       
         
           
           
               
               
           
         
       
       wherein the resulting product is filtrated to remove the organic solvent and then washed by an organic solvent; and Fmoc is a protecting group fluorenylmethoxycarbonyl;
 (b) reacting a single-stranded DNA (ssDNA) with a second linker 
 
       
         
           
           
               
               
           
         
       
       to obtain 
       
         
           
           
               
               
           
         
       
       wherein the resulting product is precipitated, washed with ethanol, and centrifuged;
 (c) reacting 
 
       
         
           
           
               
               
           
         
       
       with 
       
         
           
           
               
               
           
         
       
       in an organic solvent to obtain 
       
         
           
           
               
               
           
         
       
       wherein the resulting product is filtrated to remove the solvent and then washed with water and a triethylammonium acetate (TEAA) buffer solution;
 (d) removing the Fmoc protecting group by contact with a base to obtain 
 
       
         
           
           
               
               
           
         
         (e) reacting with a skeleton molecule comprising a carboxylic acid which reacts with the free amine of the first linker, in an organic solvent, wherein the skeleton molecule is selected from the group consisting of 4-aminobenzoic acid, dl-4-hydroxyphenylglycine, Fmoc-glycine, Fmoc-1-phenylalanine, t-butylisocynide, cyclohexyl isocyanide, 3-methyl butyraldehyde, cyclopentyl aldehyde and 
       
       
         
           
           
               
               
           
         
         (f) reacting the product obtained in step (e) with a synthetic building block, wherein the synthetic building block is selected from the group consisting of isocyanate, benzyl alcohol and benzoic acid; 
         (g) perform DNA-encoding on the product obtained in step (f), wherein the synthetic building block is specifically labeled by a marker sequence which is connected to the ssDNA; and 
         (h) cleaving the product obtained in step (g) from the solid carrier to obtain a DNA-encoded compound library. 
       
     
     
       2. The method of  claim 1 , wherein in step (a), the organic solvent for reaction is dichloromethane. 
     
     
       3. The method of  claim 1 , wherein the product obtained in step (a) is washed by DMF and dichloromethane. 
     
     
       4. The method of  claim 1 , wherein in step (d), the base is pyridine. 
     
     
       5. The method of  claim 1 , wherein the ssDNA has the sequence of GGAGCTTGTGAATTCTGGCACTCG. 
     
     
       6. The method of  claim 1 , further comprising: before step (f), performing DNA-encoding on the product obtained in (e), wherein the skeleton molecule is specifically labeled by a marker sequence which is connected to the ssDNA. 
     
     
       7. The method of  claim 1 , wherein in step (h), the step of cleaving the product obtained in step (g) from the solid carrier is carried out in ammonium hydroxide. 
     
     
       8. The method of  claim 1 , wherein in step (e), the skeleton molecule is 4-aminobenzoic acid. 
     
     
       9. The method of  claim 8 , wherein the product obtained in step (e) is 
       
         
           
           
               
               
           
         
       
     
     
       10. The method of  claim 1 , wherein in step (e), the skeleton molecule is 
       
         
           
           
               
               
           
         
       
     
     
       11. The method of  claim 10 , wherein the product obtained in step (e) is 
       
         
           
           
               
               
           
         
       
     
     
       12. The method of  claim 1 , wherein in step (e), the skeleton molecule is a combination of Fmoc-glycine, t-butylisocynide and 3-methyl butyraldehyde. 
     
     
       13. The method of  claim 1 , wherein in step (e), the skeleton molecule is a combination of Fmoc-1-phenylalanine, cyclohexyl isocyanide and cyclopentyl aldehyde. 
     
     
       14. The method of  claim 1 , wherein in step (f), the synthetic building block is isocyanate. 
     
     
       15. The method of  claim 1 , herein in step (f), the synthetic building block is benzoic acid.